Dr Florian Buhr

Postdoctoral research associate

Since 10/2016

Vendruscolo Lab, Center for Misfolding Diseases, University of Cambridge, UK

• Establishing a new research area at the Centre:
  mRNA mistranslation in neurodegeneration
• Research focus: Ribosomal frameshifting in Huntington Disease and other polyglutamine disorders
• Since 2022: Research collaboration with Dr Jeffrey Savas, Northwestern Univesity: Development of proteomics workflows for extremely challenging samples (low-abundance, partially mistranslated species)
• Since 2020: Leader of a project team of PhD and medical students
• 2018 - 2020: Co-organiser of the biophysical seminar lecture series hosting leading scientists in neurodegeneration and related fields
• Other responsibilities: Biological risk assessment and safety training officer, user training and administration of analytical equipment

Clinical laboratory scientist

04/2020 – 08/2020

Institut für Laboratoriumsmedizin, Mikrobiologie und Krankenhaushygiene (ILMH)
Klinikum Bayreuth GmbH, Bayreuth, Germany
(Temporary position during the COVID-19 lockdown of the University of Cambridge)

• RT-qPCR diagnostics of SARS-CoV-2 from human samples
• Introduction and validation of automated liquid handling systems and optimisation of RNA extraction SOPs, training of technical staff
• Upscaling of routine testing capabilites under dynamic reagent supply conditions
• Technical and medical interpretation of test results

Research Assistant

04/2008 – 09/2016

Schwalbe Lab, Institute for Organic Chemistry and Chemical Biology
Center for Biomolecular Magnetic Resonance (BMRZ)
Goethe University Frankfurt/Main, Germany

• Research areas: Structural biology, cotranslational protein folding, codon usage, mechanisms of mRNA riboswitches, development of photoactivatable compounds as research tools for subcellular translation monitoring
• Extensive, award-winning teaching (organic chemistry, biophysical chemistry)
• Mentoring and training of PhD and undergraduate students
• Other Responsibilities: User training and administration of analytical equipment, including procurement and price negotiations

Part-time student jobs

2007 - 2010

• Product portfolio research, translation of internal company ressources
  (Roche Diagnostics, Mannheim, Germany)
• Translation of scientific journal articles
  (Angewandte Chemie, Wiley-VCH, Weinheim, Germany)
• Teaching assistant, practical organic chemistry for Biology students
  (Goethe-University Frankfurt/Main, Gerrmany)

Compulsory national service

04/2002 – 01/2003

Nordseeklinik Borkum, Germany

Department of occupational therapy

PhD in Biochemistry (magna cum laude)

04/2008 – 09/2016

Goethe University Frankfurt/Main, Germany

Thesis title: NMR Investigations on Cotranslational Protein Folding
Supervisor: Prof Harald Schwalbe
(Dr phil. nat., awarded in 2018)

Diploma in Biochemistry (with distinction, equiv. to M.Sc.)

04/2003 – 03/2008

Goethe University Frankfurt/Main, Germany

Henkel Sponsorship Award for best Biochemistry degree in 2008
Main subjects: Biochemistry, biophysical chemistry, cell biology, bioinformatics
Thesis title: NMR Investigations on Protein Folding

Extracurricular activities:

Departmental Student organization (2004 - 2006)

Frankfurt Biochemistry Association (founding member)

Faculty council 2nd elected student representative (2005)

Secondary school

03/2002

Martin-Butzer-Gymnasium Dierdorf, Germany

Early Abitur (A levels) after skipping the 9th grade
Final grade: 1.4 (grading scale 1.0 > 4.0)

Mistranslation in neurodegeneration

Buhr F, Ciryam P, Vendruscolo M. A mistranslation-prone transcriptome underlying polyglutamine expansion diseases.
Nature Reviews Molecular Cell Biology (2021) https://doi.org/10.1038/s41580-021-00368-4

• A mistranslation hypothesis of aggregation-independent toxicity in repeat expansion diseases
• We propose a disease mechanism for polyglutamine (polyQ) diseases in which the presence of trinucleotide repeat expansions puts the overall translation system under stress. This stress is tRNA-specific and results in the failure to correctly translate a vulnerable sub-transcriptome containing CAG-rich, mistranslation-prone transcripts.

Codon usage, contranslational protein folding

Buhr F, Jha S, Thommen M, Mittelstaet J, Kutz F, Schwalbe H, Rodnina M, Komar AA. Synonymous codons direct cotranslational folding toward different protein conformations. Molecular Cell 61(3):341-51 (2016)

• International collaboration with Prof Anton Komar
  (Cleveland State University, USA)
  and Prof Marina Rodnina
  (Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany)
• Key publication in an emerging field, cited over 300 times
• Key contributions:
  Central role in experimental design and interpretation of results
  Protein expression, purification, and NMR spectroscopy

Schulte L,Mao J, Reitz J, Sreeramulu S, Kudlinzki D, Hodirnau V, Meier-Credo J, Saxena K, Buhr F, Langer JD, Blackledge M, Frangakis AS, Glaubitz C, Schwalbe H. Cysteine oxidation and disulfide formation in the ribosomal exit tunnel. Nature communications 11 (1), 1-11 (2020)

• International collaboration between a number of technically specialised labs
• Disulfide formation inside the ribosomal exit tunnel shown for the first time
• Follow-up publication to aspects of my PhD thesis
• Key contributions:
  Initial DNA construct design, data interpretation, manuscript preparation

Liu Y, Sharp JS, Do DHT, Kahn RA, Schwalbe H, Buhr F, Prestegard JH. Mistakes in translation: Reflections on mechanism.
PloS one 12 (6), e0180566 (2017)

• International collaboration with Prof James Prestegard
  (University of Georgia, USA)
• Analysis of mistranslation events during protein overexpression in E. coli by mass spectrometry
• Key contributions:
  Data analysis and interpretation

Caged puromycin, translation monitoring

Buhr F, Kohl-Landgraf J, tom Dieck S, Hanus C, Chatterjee D, Hegelein A, Schuman EM, Wachtveitl J, Schwalbe H. Design of photocaged puromycin for nascent polypeptide release and spatiotemporal monitoring of translation. Angew. Chem. Int. Ed. Engl. 54(12): 3717–3721 (2015)

• Collaboration with Erin Schuman (Max-Planck-Institute for Brain Research, Frankfurt, Germany)
• Development of a unique tool for subcellular translation monitoring
• Key contributions:
  Lead in experimental design and interpretation of results
  Compound synthesis, uncaging experiments, translation and cell toxicity assays

Kohl-Landgraf J, Buhr F, Lefrancois D, Mewes JM, Schwalbe H, Dreuw A, Wachtveitl J. Mechanism of the photoinduced uncaging reaction of puromycin protected by a 6-nitroveratryloxycarbonyl group.
JACS, 136(9):3430-8 (2014)

• Detailed photochemical characterisation of a widely used photolabile protecting group
• Key contributions:
  Compound synthesis

RNA riboswitches

Reining A, Nozinovic S, Schlepckow K, Buhr F, Fürtig B, Schwalbe H. Three-state mechanism couples ligand and temperature sensing in riboswitches. Nature, 499(7458):355-9 (2013)

• Discovery of a new mechanism by which riboswitches respond to temperature and ligand concentration
• Key contributions:
  Development of a translation assay that greatly enhanced the biological significance of the structural model, allowing its publication in Nature

Popular Science

Schwalbe H, Wachtveitl J, Heckel A, Buhr F, Schimmer TM. Komplexität und Emergenz in der Chemie Moleküle der zellulären Informationsübertragung
In: Schwalbe, Lutz-Bachmann: Komplexität – System – Evolution Eine transdisziplinäre Forschungsperspektive (2022). ISBN 978-3-495-49226-0

• Book chapter about a definition of complexity in biochemical data storage and the unlikely evolution of efficient protein folding as part of a multidisciplinary collection of essays on complexity.
• Published in German

Mistranslation in neurodegeneration

Buhr F, Ciryam P, Hanna, N, Savas, J, Vendruscolo M. Ribosomal frameshifting of endogenous transcripts in polyQ diseases (working title, manuscript in preparation).

• According to the concept of the Mistranslation-Prone Transcriptome (MPT), we characterize a set of human transcripts expected to be susceptible to ribosomal frameshifting in Huntington's Disease.
• Using a fluorescence-based frameshifting assay, we demonstrate that frameshifting frequency is dependent on polyCAG length and can be modified by sub-inhibitory concentrations of anisomycin.
• We demonstrate that the expression of frameshifted versions of transcription factors within the MPT is toxic in all conditions, while polyQ toxicity requires further stress and can be mitigated by frameshifting inhibiiton.
• Using a customised proteomics workflow, we detect frameshifted species of MPT candidates in cell models of Huntington's disease.

On protein folding and disulfide formation

Since 2017

Scientific Reports (Nature Journals)

Plenary talks

5th annual CMD conference, Taormina, Italy (2018)

Translational depletion in PolyQ diseases: An alternative mechanism that does not depend on protein-protein interactions

4th annual CMD conference, Seville, Spain (2016)

Project pitch: tRNA in Neurodegeneration

Gordon Research Conference Biopolymers, Newport, RI, USA (2014)

Effects of Synonymous Codon Mutations on the Structure of Gamma-B Crystallin

Poster presentations

Gordon Research Conference Biopolymers, Newport, RI, USA (2014)

Effects of Synonymous Codon Mutations on the Structure of Gamma-B Crystallin

Gordon Research Conference Biopolymers, Newport, RI, USA (2012)

Photocaged Puromycin for Subcellular Translation Monitoring

Euromar, Frankfurt, Germany (2011)

Triggered Polypeptide Release in Kinetic NMR experiments
on Ribosome-Nascent Chain Complexes

International Conference on Magnetic Resonance in Biological Systems (ICMRBS), Cairns, Australia (2010)

Fast NMR experiments on Ribosome-Nascent Chain Complexes

Main scientific expertise

Neurodegenerative disorders

Focus on Huntington's disease, Spinocerebellar ataxia, and Alzheimer's disease

Novel toxicity mechanisms

RNA

Structural biology

mRNA translation

Posttranscriptional regulation

Mistranslation, ribosomal frameshifting

Codon usage, tRNA pools

Protein folding

Structural biology

Misfolding and aggregation

Cotranslational folding

Co- and posttranslational modifications

Protein homeostasis

Technical Skills

Proteomics

LC-MS/MS sample preparation from cultured cells and tissues

Subcellular fractionation (nuclei, ribosomes, solubility)

IP-based sample preparation

Translatomics (Punch-P)

Custom database design

Data analysis (MaxQuant)

Analytical protein biochemistry

PAGE (SDS, native, IEF)

Western Blotting, ELISA

Immunoprecipitation (IP, Co-IP)

Assay development

in vitro luciferase translation assay for riboswitch mRNA regulation

DNA template design, readout optimisation

Temperature- and ligand screening

Ribosomal frameshifting assays

Cell viability/toxicity assays

Biophysical methods

Optical spectroscopy (UV/Vis, IR, CD, fluorescence)

Stopped-flow kinetics

Microscale thermophoresis (MST)

NMR Spectroscopy

All aspects of sample preparation and experimental setup (Bruker)

2D/3D Protein NMR (including resonance assignment)

Fast/kinetic 2D NMR, laser-coupled 1D experiments

NMR on challenging samples (molecular weight)

HRMAS, metabolomics

Molecular Biology

Standard cloning techniques (PCR, restriction, ligation, transformation)

Design of fusion proteins, affinity tags, linkers, ribosome stalling sequences

Codon optimisation (codon harmonisation)

Site-directed mutagenesis, TOPO-cloning

Cloning of difficult constructs (riboswitches, polyCAG tracts)

Synthetic Organic Chemistry

Basic preparative and analytical techniques

Synthesis of photolabile protecting groups

RP-HPLC

Cell culture

Cell lines:

Human: SH-SY5Y, HEK293/HEK293T, HeLa

Rodent: N2a, PC12

IPSC-derived: iCell GABANeurons

Insect: High 5/BTI-Tn-5B1-4, SF9

Techniques:

Adherent and suspension culture

Transfection: Lipofection, electroporation, magnetic beads

Generation of stable transfectant cell lines

Differentiation: Dopaminergic neurons

Fluorescence microscopy, flow cytometry, FACS

Stress condition screens, toxicity assays

Protein expression and purification

Protein expression

Expression systems:

E. coli

P. pastoris

Insect cells

cell-free / in vitro protein expression

Special techniques:

Coexpression of chaperones or rare tRNA species

Selective isotope-labelling (amino acid-specific, pulse-labelling)

Expression of stalled ribosome-nascent chain complexes

Expression under non-reducing / oxidising conditions

Protein purification

Affinity chromatography (IMAC, Strep-tag, FLAG-tag)

FPLC (IEC/IEX, SEC)

Preparative IP

RT-qPCR

RNA extraction from human samples (nasopharyngeal swabs, biosafety level 2)

Setup and operation of automated liquid handling platforms (Hamilton)

RT-qPCR and data analysis

Software Skills

Programming

Python (intermediate)

RStudio (beginner)

Web Development

HTML, CSS, Django (intermediate)

Data analysis software

NMR (Bruker TopSpin)

Proteomics (MaxQuant)

Imaging (FIJI/ImageJ)

Flow Cytometry (FlowJo)

Python packages (FlowCal, SciPy, Pandas)

Office and Visualization

MS-Office (Word, Excel, Powerpoint, advanced)

Adobe Creative Suite (Illustrator, Photoshop, advanced)

Python packages (Seaborn, Matplotlib)

Miscellaneous

Cloning (Genome Compiler)

Structural Biology (PyMol, Chimera)

Instrument-specific packages

Video editing, 3D printing & CAD design (basic)

Postgraduate

Since 2014

Mentoring and training of PhD students

• Introduction to key literature in the field
• Practical training and research strategy

Undergraduate

Since 2008

Supervision of master’s theses and undergraduate research students

2008 - 2012

Development and teaching of undergraduate seminars in Organic Chemistry for Chemistry students

• Departmental award for outstanding teaching

2005 - 2015

Teaching assistant in undergraduate practical courses

• Organic Chemistry
• Biophysics

Invited Talks (selected)

Since 2015

Presentations and lectures on recurring topics
(Alzheimer’s disease, RNA, food science)

• Leibniz-Institute for Food Systems Biology, TU Munich, Germany (2018)
• Royal Society of Chemistry, London, UK (2017)
• Diet Body Brain Competence Cluster, University of Bonn, Germany (2017)
• Rotary Club Frankfurt am Main - International (2015)

Since 2013

Science Slam presentations in 20+ German cities and online

• Competitive 10 minute presentations in front of a live audience
• Scientific infotainment, 4-6 participants
• Placement rate: First: >50%, First or Second: >75%
• Video examples here and here (in German)
• Press releases here, here and here (in German)

Since 2009

Night of Science, Goethe University Frankfurt/Main, Germany

• Annual lectures and experimental demonstrations
• 2020 online lecture mirrored here (in German)

Teaching

2009

Departmental award for outstanding teaching
Department of Chemistry, Goethe University Frankfurt/Main, Germany

• First time ever awarded to a PhD student

Academic

2009

Invited participant of the 59th Meeting of Nobel Laureates at Lake Constance,
Lindau, Germany

2008

Henkel Sponsorship Award for best Biochemistry degree in 2008
Goethe University Frankfurt/Main, Germany

Scholarships

2008

Graduate Scholarship, German Chemical Industry Fund

2005

Undergraduate scholarship, German National Academic Foundation

Regulatory

2016

GMP and Quality Management in the Pharmaceutical Industry, (two-day introductory course), Goethe-University, Fankfurt, Germany

Technical

2010

ÄKTA workshop protein chromatography, GE Healthcare, Munich, Germany

Advanced course in multidimensional NMR in solution,
James Cook University, Cairns, Australia

Toxicology

2008

Certificate of competence according to § 5 section 2 ChemVerbotsV

Research Collaborations

2011

Prof Marina Rodina, Max-Planck-Institute for Biophysical Chemistry,
Göttingen, Germany

• Preparation of NMR samples with advanced isotope labelling schemes
• Preparation of ribosomal complexes

2009

Prof John Christodoulou, University College London, UK

• In vitro protein synthesis

Undergraduate Internships

2006

Prof Wendell Lim, University of California, San Francisco, CA, USA

• Research topic: Signal transduction
• Protein expression and purification
• Protein crystallography

2005

Prof Holger Gohlke, Goethe University Frankfurt/Main, Germany

• Research topic: Parametrisation of kinase structures
• Structural biology, bioinformatics, C++ programming

2004

Prof Hartmut Michel, Max-Planck-Institute for Biophysics, Frankfurt/Main, Germany

• Research topic: Use of antibody fragments as stabilisers of membrane proteins in protein crystallography
• Protein expression and purification

Since 2017

DKMS UK

Founder and leader of the Cambridge volunteer hub

• Registered 1500+ new stem cell donors

2009 - 2011

Local Fundraising

Community youth projects, Frankfurt, Germany

Language Proficiency

German: Native speaker

English: Full professional (CEFR C2/ILR 4+)

Driving licence

European class B

Personal Interests

Science communication, food science, board games